G-protein signaling abnormalities mediated by CD95 in salivary epithelial cells.

Salivary epithelial cells from patients with primary Sjögren's syndrome (SS) undergo Fas-mediated apoptosis. Bcl-2 and Bcl-xL are apoptosis suppressing oncogenes. Very little is known about the role of these oncogene molecules in salivary epithelial cells. To investigate the possible prevention of salivary glandular destruction in SS by Bcl-2 and Bcl-xL, stable transfectants expressing these molecules were made from HSY cells, a human salivary epithelial cell line. HSY cells were transfected with an expression vector for human Bcl-2 or Bcl-xL. Stable transfectants were selected and apoptosis was induced by anti-Fas antibody. Apoptosis was quantified by propidium iodide staining followed by flow cytometry. Caspase activity was detected by immunohistochemical analysis and enzyme cleavage of DEVD-AMC, a fluorescent substrate. Response to carbachol, a muscarinic receptor agonist, and EGF was measured by Ca2+ mobilization and influx. Fas-mediated apoptosis was significantly inhibited in Bcl-2 and Bcl-xL transfectants compared to wild-type and control transfectants (empty vector). Surprisingly, caspase activity was not inhibited in Bcl-2 and Bcl-xL transfectants. Activation of the Fas pathway in the Bcl-2 and Bcl-xL transfectants by antibody also inhibited carbachol and EGF responsiveness (i.e., Ca2+ mobilization and/or influx) by 50-60%. This Fas-mediated inhibition of cell activation was partially or completely restored by specific peptide interference of caspase enzyme activity. The prevention of Fas-mediated apoptosis by the overexpression of Bcl-2 and Bcl-xL in salivary gland epithelial cells results in injured cells expressing caspase activity and unable to respond normally to receptor agonists. Such damaged cells may exist in SS patients and could explain the severe dryness out of proportion to the actual number of apoptotic cells seen on salivary gland biopsy.

Fas-mediated apoptosis in a rat acinar cell line is dependent on caspase-1 activity.

Apoptosis plays an important role in the dysfunction of exocrine glands. Fas is a death-inducing receptor found on many types of cells including epithelial acinar cells. To elucidate the intracellular mechanism of Fas-mediated cell death in exocrine glands, an epithelial acinar cell line, SMG-C6, was studied. Caspase-1, -3, -8, and -9 activities were elevated in SMG-C6 cells after the induction of apoptosis by soluble Fas ligand (FasL). The activation of caspase-1 and -8 occurred prior to caspase-3 and -9 activation. The caspase-1 inhibitor, zYVAD-fmk, was effective in preventing cell death, whereas the caspase-3 and -8 inhibitors (ac-DEVD-CHO and ac-IETD-CHO, respectively) were not. zYVAD-fmk was able to inhibit caspase-3 activation indicating that caspase-1 is upstream to caspase-3. Furthermore, kinetic studies show that caspase-1 is an early event in the Fas apoptotic pathway. This study shows that caspase-1 participates in Fas-mediated apoptosis of epithelial cells by initiating the caspase cascade.

Rheumatoid arthritis and simultaneous aortic, mitral, and tricuspid valve incompetence.

We describe a 72-year-old woman with aortic, mitral, and tricuspid valve incompetence secondary to a rheumatoid granulomata. The cardiac valvular lesions developed simultaneously and deteriorated rapidly. The patient died after a transient relief of symptoms by high dose steroid therapy.

[Cold-induced reversible brain ischemia in mixed connective tissue disease, a case report].

A 19-year-old woman with long-standing mixed connective tissue disease was admitted for dizziness. We examined cerebral blood flow quantitation using 99mTc-hexamethylpropyleneamine oxime (HMPAO) and single photon emission computed tomography (SPECT) at rest and after cold pressor test. Mean cerebral blood flow reduced remarkably when she complained dizziness and showed peripheral Raynaud's phenomenon after cold exposure. We concluded cold-induced reversible brain ischemia was the reason of dizziness. Our finding suggests brain Raynaud's phenomenon. Further studies are necessary to clarify this phenomenon.

[Cytomegalovirus infection associated with immunosuppressive therapy in collagen vascular diseases].

OBJECTIVE: We investigated the frequency of cytomegalovirus (CMV) infection during immunosuppressive therapy in collagen vascular disease by using CMV antigenemia assay. METHODS: CMV antigenemia in fifteen patients with collagen vascular disease were analyzed before and one month after immunosuppressive therapy with more than 30 mg/day of prednisolone. RESULTS: CMV antigenemia were detected in 9 patients (60%), however no CMV antigenemia detected in all patients before treatment. In 7 of 9 patients CMV infection occurred such as fever, leucocytopenia, thrombocytopenia, liver injury, interstitial pneumonia. In 2 patients of polymyositis/dermatomyositis, creatine phosphokinase (CPK) levels which had decreased once just after treatment started, elevated again although initial dose of prednisolone continued. After ganciclovir administration because of the positive results of CMV antigenemia, elevated CPK levels were normalized immediately. CONCLUSION: Our results showed that CMV infection occurred with high frequency during immunosuppressive therapy, and might mimic the exacerbation of collagen vascular disease. It is important to differentiate CMV infection from increased activity of collagen vascular disease during the treatment.

The effects of hedonic properties of odors and attentional modulation on the olfactory event-related potentials.

The purpose of this study was to investigate the influence of the hedonic properties of odors and the attention of subjects on components of the olfactory event-related potentials (OERP). The subjects were seven healthy male students. Two odors (orange and eugenol) of different hedonic properties were presented to the subjects via a constant-flow olfactometer during an oddball paradigm under ignore and attend conditions, and the OERP were then established. The latencies of the OERP were not affected by the qualitatively different odors, whereas the amplitude of late positive component (P3) during the presentation of orange was significantly larger than that during the presentation of eugenol. On the other hand, the allocation of a subject's attention led to a decrease in the latency and to an increase in the amplitude of P3. Moreover, the amplitude of P3 increased significantly when the pleasant odor (orange) in the rare stimulus was presented under the attend condition. These results suggested that hedonic property, distribution of attention, and the interaction between these factors may influence the OERP components.

Effects of inhalation of essential oils on EEG activity and sensory evaluation.

The purpose of this study was to investigate EEG changes in subjects directly after inhalation of essential oils, and subsequently, to observe any effect on subjective evaluations. EEG and sensory evaluation were assessed in 13 healthy female subjects in four odor conditions. Four odor conditions (including lavender, chamomile, sandalwood and eugenol) were applied respectively for each subject in the experiment. The results were as follows. 1) Four basic factors were extracted from 22 adjective pairs by factor analysis of the sensory evaluation. The first factor was "comfortable feeling", the second "cheerful feeling", the third "natural feeling" and the fourth "feminine feeling". In the score of the first factor (comfortable feeling), the odors in order of high contribution are lavender, eugenol, chamomile and sandalwood. 2) Alpha 1 (8-10 Hz) of EEG at parietal and posterior temporal regions significantly decreased soon after the onset of inhalation of lavender oil (p < 0.01). Significant changes of alpha 1 were also observed after inhalation of eugenol or chamomile. The change after inhalation of sandalwood was not significant. These results showed that alpha 1 activity significantly decreased under odor conditions in which subjects felt comfortable, and showed no significant change under odor conditions in which subjects felt uncomfortable. These results suggest a possible correlation between alpha 1 activity and subjective evaluation.

Bcl-2 family expression in salivary glands from patients with primary Sjögren's syndrome: involvement of Bax in salivary gland destruction.

To investigate the molecular mechanism of glandular parenchyma destruction in Sjögren's syndrome (SS), Bcl-2, Bax, Bcl-X, and Bak expression were studied. SS (n = 18) and control salivary glands (n = 6) were examined by immunohistochemistry. Apoptosis was assessed by in situ DNA nick end labeling. Infiltrating mononuclear cells in the SS salivary gland showed elevated Bcl-2. These mononuclear cells expressed increased Bax but did not undergo apoptosis. Both SS and control salivary gland ductal epithelial cells expressed Bcl-2, Bax, and Bcl-X. SS, but not normal, salivary gland acinar cells expressed Bax and underwent apoptosis. These results suggest that elevated Bax expression in SS salivary gland acinar cells may play an important role in the apoptotic pathway. In contrast, Bcl-2 expression in SS infiltrating mononuclear cells and ductal cells may contribute to their survival.

Elevated proapoptotic Bax and caspase 3 activation in the NOD.scid model of Sjögren's syndrome.

OBJECTIVE: Salivary gland epithelial cells in patients with Sjögren's syndrome (SS) and in NOD and NODscid mice express Fas and Fas ligand, and these cells die from apoptosis. To elucidate the intracellular molecular mechanisms responsible for this salivary gland epithelial cell apoptosis, expression of the Bcl-2 family of proteins (Bcl-2, Bcl-xL, Bax) and caspase (caspases 3 and 8) was studied in young (ages 8-10 weeks) and old (ages 17-28 weeks) NOD and NOD.scid mice. METHODS: Sections of frozen salivary gland tissue were obtained from NOD and NOD.scid mice and from the lip biopsy material of SS patients. Immunohistochemistry or Western blot analysis was performed to assess the apoptotic-associated proteins. RESULTS: Levels of Bax and caspase 3 were elevated in the epithelial cells of glands from old NOD mice, but not in those from young NOD mice. In contrast, epithelial cells from both young and old NOD.scid mice exhibited strong expression of Bax and caspase 3. Western blot analysis showed that the activated form of caspase 3 was increased 2-5-fold in the glands from old NOD, old NOD.scid, and young NOD.scid mice compared with those from young NOD mice. Caspase 3 was also significantly elevated (P < 0.01) in SS patients whose focus scores were grade 3 or 4. In the SS patients' biopsy tissue and in the mouse glands, cells with fragmented DNA were positive for caspase 3. CONCLUSION: These results demonstrate that salivary gland epithelial cells in NOD and NOD.scid mice overexpress the proapoptotic molecules Bax and caspase 3. Bax could be the gene responsible for initiation of caspase activation, epithelial cell destruction, and lymphocyte glandular localization in SS.